Selection of carbohydrate-binding cell phenotypes using oligosaccharide-coated magnetic particles

Abstract

Neoglycoconjugate coated magnetic beads were assessed for their ability to selectively isolate human cells with known anti-carbohydrate reactivity. Four lung cancer cell lines, NCI-H146; NCI-N417D, SKMES-1, EKVX; two acute lymphoblastic leukemia lines, MOLT-4 and CCRF-CEM; and the anti-Le(c) (isolactosamine) hybridoma, LU-BCRU-G7, were tested. The neoglycoconjugates (biotinylated pseudopolysaccharides) bound uniformly to streptavidin coated magnetic beads as demonstrated by FITC labeled lectin. Streptavidin beads alone did not bind to any of the cell types. The anti-Le(c) hybridoma cell line, LU BCRU-G7, demonstrated binding only to Le(c) pseudopolysaccharide coated magnetic beads. Subsequent incubation in the presence of unlabeled pseudopolysaccharide resulted in the release of the beads from the cell surface. Although there was some heterogeneity within the individual lung and leukemic cell lines, positive cells showed strong rosette formation with the coated beads. The A(di) disaccharide coated beads showed binding in all four lung cancer cell lines, with the Le(c) and the H (type1) pseudopolysaccharide-bead conjugates only reactive in the N417 and H146 SCLC lines. The range of L-selectin ligand-coated beads were all successful in binding to the acute lymphoblastic leukemia cell lines MOLT4 and CCRF-CEM. This approach provides a versatile model for the study of cell-surface carbohydrate interactions that should find application in many areas of cell biology.